Direct Zol RNA Miniprep Kit Manual

The Direct Zol RNA Miniprep Kit is designed for rapid purification of high-quality total RNA from various sample types including cells, tissues, and liquid samples. Utilizing Zol reagent for efficient lysis and RNA stabilization, this kit provides a streamlined workflow with spin column technology, DNase treatment capability, and elution in RNase-free water. Below are key sections for product overview, components, protocol, applications, troubleshooting, and storage information.

1. Kit Components 2. Storage Conditions 3. Protocol Overview 4. Detailed Procedure 5. Expected Results 6. Applications 7. Troubleshooting 8. Safety Information 9. Technical Support 10. Ordering Information

Kit Components

Each Direct Zol RNA Miniprep Kit contains sufficient reagents for 50 purifications.

Component	Description	Quantity
Zol Reagent	Complete lysis solution for RNA stabilization	100 ml
Spin Columns	RNA-binding silica membrane columns	50 columns
Collection Tubes	2 ml tubes for centrifugation	50 tubes
Wash Buffer 1	First wash solution	40 ml
Wash Buffer 2	Second wash solution (concentrated)	15 ml
DNase I	Lyophilized DNase for genomic DNA removal	500 units
DNase Reaction Buffer	Buffer for DNase treatment	1.5 ml
RNase-free Water	Elution solution	10 ml

Storage Conditions

Proper storage ensures reagent stability and performance.

Zol Reagent: Store at 4°C; stable for 12 months
Wash Buffers: Store at room temperature; add ethanol before use
DNase I: Store at -20°C; reconstitute before use
Spin Columns: Store at room temperature
RNase-free Water: Store at room temperature

NOTE: All components are stable until expiration date when stored as directed.

Protocol Overview

Complete RNA purification in less than 30 minutes.

Sample homogenization in Zol Reagent
Centrifugation to remove insoluble material
RNA binding to spin column
DNase treatment (optional)
Wash steps
RNA elution in RNase-free water

CAUTION! Work in RNase-free environment; use appropriate personal protective equipment.

Detailed Procedure

Follow these steps for optimal RNA yield and quality.

Homogenization: Add 100-500 mg tissue or 10^6 cells to 1 ml Zol Reagent; homogenize thoroughly
Incubation: Incubate 5 minutes at room temperature
Centrifugation: Centrifuge at 12,000 × g for 5 minutes at 4°C
Binding: Transfer supernatant to spin column; centrifuge 30 seconds
DNase Treatment: Add DNase mix directly to membrane; incubate 15 minutes
Wash 1: Add 700 μl Wash Buffer 1; centrifuge 30 seconds
Wash 2: Add 500 μl Wash Buffer 2; centrifuge 30 seconds
Elution: Add 30-50 μl RNase-free water; centrifuge 1 minute

Expected Results

Typical performance characteristics of the Direct Zol RNA Miniprep Kit.

Yield: 5-30 μg total RNA per purification
Purity: A260/A280 ratio of 1.8-2.1
RNA Integrity: RIN values typically >8.0
DNA Contamination: Minimal genomic DNA carryover
Applications: Suitable for RT-PCR, qPCR, microarray, sequencing

Applications

The purified RNA is compatible with various downstream applications.

Application	Compatibility	Notes
RT-PCR	Excellent	No additional purification required
qPCR	Excellent	Consistent Ct values
RNA Sequencing	Good	High-quality libraries
Microarray	Good	Consistent hybridization
Northern Blot	Good	Intact RNA bands

Troubleshooting

Common issues and solutions for RNA purification.

Problem	Possible Cause	Solution
Low yield	Incomplete homogenization	Ensure thorough sample disruption
Poor A260/A280	Carryover of Zol Reagent	Ensure complete washing
DNA contamination	DNase not effective	Check DNase activity and incubation time
Degraded RNA	RNase contamination	Use RNase-free techniques and reagents
Clogged column	Excess cellular debris	Centrifuge longer after homogenization

Safety Information

Zol Reagent contains guanidine salts and phenol. Handle with care.

Wear gloves, lab coat, and safety glasses
Work in well-ventilated area
Avoid contact with skin and eyes
Dispose of waste according to local regulations
In case of contact, flush with plenty of water

WARNING! Zol Reagent is toxic if ingested and causes skin irritation.

Technical Support

For technical assistance and product inquiries.

Email: [email protected]
Phone: 1-800-123-4567
Website: www.company.com/support
Hours: Monday-Friday, 8:00 AM - 5:00 PM EST

Include product lot number and detailed description of issue when contacting support.

Ordering Information

Product specifications and ordering details.

Catalog Number	Description	Purifications	Price
DZ-R50	Direct Zol RNA Miniprep Kit	50	$250.00
DZ-R100	Direct Zol RNA Miniprep Kit	100	$450.00
DZ-R250	Direct Zol RNA Miniprep Kit	250	$950.00

Note: Prices subject to change; contact for bulk pricing.